胞红蛋白在缺氧神经元中的表达

Expression of cytoglobin in hypoxic neurons in vitro

目的探讨细胞红蛋白（cytoglobin,CYGB）在体外培养神经元缺氧时的表达,为研究缺氧缺血性脑病（hypoxic ischemic encephalopathy,HIE）的生理病理机制及寻求该病新的治疗方法提供实验基础。方法运用无血清培养技术进行胎鼠皮质神经元原代培养6d;先用神经元特异性烯醇化酶免疫组织化学方法纯度鉴定,然后随机分配为4组,将其中3组（第1组作为对照）置于0.5%-0.9%的O2中,用CCK8试剂盒测试细胞生存率,同时用Real-timePCR和Westernblot观察CYGB在神经元正常及缺氧不同时间点（8h,16h,24h）核酸及蛋白的表达情况。结果神经元纯度鉴定纯度大于99%;缺氧后神经元生存率随着时间推移降低,但绝大部分神经元仍存活;CYGB核酸及蛋白的表达随缺氧时间增长而递增,且每组之间差异有统计学意义（P〈0.05）。结论在体外培养的神经元细胞中,缺氧可以促进CYGB的表达,提示CYGB在缺氧性神经损伤中可能发挥某种特定的生理功能。

Objective To examine the expression of eytoglobin（CYGB） of neurons in vitro when suffered hypoxia,which provides an experimental basis for exploring the physiological and pathological mechanism and seeking new treatmentsof hypoxic-ischemic encephalopathy, methods The primary cortical neurons from embryonic rat were cultured in a serum-free culture system of B27-supplemented neurobasal medium. The neurons were identified by polyclonal antibody againstneuron specific enolase（NSE） on the 6th day. They were divided into 4 groups randomly for assessing cell viability by CCK8and observing the expression of CYGB by Real-time PCR and Westernblot at 0,8,16,24 h after hypoxic model（oxygen con-centration between 0.5%and 0.9 %） was induced, results The purity of neurons was greater than 99% ;and cell viabilitywere slightly reduced after hypoxia. Expression of RNA and protein of CYGB was increased in a time-dependent manner.And there was statistical, differenc in each group （P〈0.05）. conclusions CYGI3 can up regulate in neurons in vitro afterhypoxia,indicating CYGB may play certain important roles in neurons under hypoxia.